Nitric oxide regulates oxygen uptake and hydrogen peroxide release by the isolated beating rat heart.
Isolated rat heart perfused with 1.5-7.5 microM NO solutions or bradykinin, which activates endothelial NO synthase, showed a dose-dependent decrease in myocardial O2 uptake from 3.2 +/- 0.3 to 1.6 +/- 0.1 (7.5 microM NO, n = 18, P < 0.05) and to 1.2 +/- 0.1 microM O2.min-1.g tissue-1 (10 microM bradykinin, n = 10, P < 0.05). Perfused NO concentrations correlated with an induced release of hydrogen peroxide (H2O2) in the effluent (r = 0.99, P < 0.01). NO markedly decreased the O2 uptake of isolated rat heart mitochondria (50% inhibition at 0.4 microM NO, r = 0.99, P < 0.001). Cytochrome spectra in NO-treated submitochondrial particles showed a double inhibition of electron transfer at cytochrome oxidase and between cytochrome b and cytochrome c, which accounts for the effects in O2 uptake and H2O2 release. Most NO was bound to myoglobin; this fact is consistent with NO steady-state concentrations of 0.1-0.3 microM, which affect mitochondria. In the intact heart, finely adjusted NO concentrations regulate mitochondrial O2 uptake and superoxide anion production (reflected by H2O2), which in turn contributes to the physiological clearance of NO through peroxynitrite formation.
Poderoso
J J
JJ
Laboratory of Oxygen Metabolism, University Hospital, School of Medicine, Buenos Aires, Argentina.
Peralta
J G
JG
Lisdero
C L
CL
Carreras
M C
MC
Radisic
M
M
Schöpfer
F
F
Cadenas
E
E
Boveris
A
A
D000818:Animals; D001920:Bradykinin / Q000494:pharmacology*; D003574:Cytochrome c Group / Q000378:metabolism; D004227:Dithionite / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000378:metabolism; D005260:Female; D006321:Heart / Q000187:drug effects / Q000502:physiology*; D006861:Hydrogen Peroxide / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D009200:Myocardial Contraction; D009569:Nitric Oxide / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D010101:Oxygen Consumption / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D003574:Cytochrome c Group; D004227:Dithionite; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D019001:Nitric Oxide Synthase; D003576:Electron Transport Complex IV; D001920:Bradykinin